Vasques Enio Rodrigues, Cunha Jose Eduardo Monteiro, Coelho Ana Maria Mendonca, Sampietre Sandra N, Patzina Rosely Antunes, Abdo Emilio Elias, Nader Helena B, Tersariol Ivarne L S, Lima Marcelo Andrade, Godoy Carlos M G, Rodrigues Tiago, Chaib Eleazar, D'Albuquerque Luiz A C
Department of Gastroenterology (LIM 37), Medical School, University of Sao Paulo (USP), Sao Paulo, Brazil.
Department of Biochemistry, Federal University of Sao Paulo (UNIFESP), Sao Paulo, Brazil.
PLoS One. 2016 Feb 22;11(2):e0149630. doi: 10.1371/journal.pone.0149630. eCollection 2016.
Ischemia and reperfusion (I/R) causes tissue damage and intracellular calcium levels are a factor of cell death. Sodium calcium exchanger (NCX) regulates calcium extrusion and Trisulfated Disaccharide (TD) acts on NCX decreasing intracellular calcium through the inhibition of the exchange inhibitory peptide (XIP).
The aims of this research are to evaluate TD effects in liver injury secondary to I/R in animals and in vitro action on cytosolic calcium of hepatocytes cultures under calcium overload.
Wistar rats submitted to partial liver ischemia were divided in groups:
(n = 10): surgical manipulation with no liver ischemia; Saline: (n = 15): rats receiving IV saline before reperfusion; and TD: (n = 15): rats receiving IV TD before reperfusion. Four hours after reperfusion, serum levels of AST, ALT, TNF-α, IL-6, and IL-10 were measured. Liver tissue samples were collected for mitochondrial function and malondialdehyde (MDA) content. Pulmonary vascular permeability and histologic parameters of liver were determined. TD effect on cytosolic calcium was evaluated in BRL3A hepatic rat cell cultures stimulated by thapsigargin pre and after treatment with TD.
AST, ALT, cytokines, liver MDA, mitochondrial dysfunction and hepatic histologic injury scores were less in TD group when compared to Saline Group (p<0.05) with no differences in pulmonary vascular permeability. In culture cells, TD diminished the intracellular calcium raise and prevented the calcium increase pre and after treatment with thapsigargin, respectively.
TD decreases liver cell damage, preserves mitochondrial function and increases hepatic tolerance to I/R injury by calcium extrusion in Ca2+ overload situations.
缺血再灌注（I/R）会导致组织损伤，细胞内钙水平是细胞死亡的一个因素。钠钙交换体（NCX）调节钙的排出，三硫酸化二糖（TD）作用于NCX，通过抑制交换抑制肽（XIP）来降低细胞内钙水平。
本研究旨在评估TD对动物I/R继发性肝损伤的影响以及在钙超载情况下对肝细胞培养物胞质钙的体外作用。
将接受部分肝脏缺血的Wistar大鼠分为以下几组：
（n = 10）：进行无肝脏缺血的手术操作；
（n = 15）：在再灌注前接受静脉注射生理盐水的大鼠；
TD组：（n = 15）：在再灌注前接受静脉注射TD的大鼠。再灌注4小时后，测量血清中AST、ALT、TNF-α、IL-6和IL-10的水平。收集肝脏组织样本以检测线粒体功能和丙二醛（MDA）含量。测定肺血管通透性和肝脏的组织学参数。在毒胡萝卜素刺激前后，评估TD对BRL3A大鼠肝细胞培养物胞质钙的影响。
与生理盐水组相比，TD组的AST、ALT、细胞因子、肝脏MDA、线粒体功能障碍和肝脏组织学损伤评分更低（p<0.05），肺血管通透性无差异。在培养细胞中，TD减少了细胞内钙的升高，并分别在毒胡萝卜素处理前后防止了钙的增加。
在Ca2+超载情况下，TD通过钙排出减少肝细胞损伤，保留线粒体功能，并增加肝脏对I/R损伤的耐受性。
